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Be certain that all of the purification buffers are at 4 °C and perform the protein purification in the chilly room to avoid protein denaturation. NOTICE: This avoids the coupling of amino acids in the further cycles with the membrane instead of the rising peptide chain. NOTE: After the coupling of the first C-terminal amino acid of all peptides to the amino group of the membrane, the amino teams between the spots and likewise a number of the amino groups inside the spot areas don't kind bonds with the amino acids.
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